| 產品名稱 |
T47D-KBluc |
| 商品貨號 |
B238518 |
| Organism |
Homo sapiens, human |
| Tissue |
mammary gland; breast/duct; derived from metastatic site: pleural effusion |
| Cell Type |
epithelial transfected with reporter plasmid |
| Product Format |
frozen |
| Morphology |
epithelial |
| Culture Properties |
adherent |
| Biosafety Level |
1
Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country. |
| Disease |
ductal carcinoma |
| Age |
54 years |
| Gender |
female |
| Applications |
They can be used to screen chemicals for estrogenic or anti-estrogenic activity. |
| Storage Conditions |
liquid nitrogen vapor phase |
| Clinical Data |
female |
| HeLa Markers |
N |
| Comments |
The parent cell line was transfected with pGL2.TATA.Inr.luc.neo which contains three estrogen responsive elements upstream of a luc reporter gene.
The cells were selected for responsiveness to 17-beta-estradiol. They can be used to screen chemicals for estrogenic or anti-estrogenic activity. |
| Complete Growth Medium |
The base medium for this cell line is ATCC-formulated RPMI-1640 Medium, Catalog No. 30-2001. To make the complete growth medium, add the following components to the base medium: 0.2 Units/ml bovine insulin; fetal bovine serum to a final concentration of 10%.
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| Subculturing |
Volumes used in this protocol are for 75cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.
Note: Subculture cells before or upon reaching confluence. Do not allow cells to become super-confluent.
- Remove and discard culture medium.
- Briefly rinse the cell layer with either Hanks' Balanced Salt Solution (HBSS) or 0.25% (w/v) Trypsin - 0.53 mM EDTA solution to remove all traces of serum that contains trypsin inhibitor.
- Add 1.0 to 2.0 mL of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes).
Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37°C to facilitate dispersal.
- Add 10 to 15 mL of complete growth medium and aspirate cells by gently pipetting.
- Add appropriate aliquots of the cell suspension to new culture vessels. An inoculum of 2 x 104 to 4 x 104 viable cells/cm2 is recommended.
- Incubate cultures at 37C. We recommend that you maintain cultures at a cell concentration between 3 x 104 and 2 x 105 cells/cm2.
Subcultivation Ratio: 1:2 to 1:3 is recommended
Medium Renewal: every 2 to 3 days
Note: The depositor states that supplementing the growth medium with insulin is optional. |
| Cryopreservation |
Freeze medium: culture medium, 95%; DMSO, 5% Storage temperature: liquid nitrogen vapor phase |
| Culture Conditions |
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37°C |
| STR Profile |
Amelogenin: X CSF1PO: 11,13 D13S317: 12 D16S539: 10 D5S818: 12 D7S820: 11 THO1: 6 TPOX: 11 vWA: 14 |
| Population Doubling Time |
36 |
| Name of Depositor |
VS Wilson |
| Year of Origin |
January 4, 2000 |
| References |
Wilson VS, et al. Development and characterization of a cell line that stably expresses an estrogen-responsive luciferase reporter for the detection of estrogen receptor agonist and antagonists. Toxicol. Sci. 81: 69-77, 2004. PubMed: 15166400
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